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Phenotypic alteration of eukaryotic cells using randomized libraries of artificial transcription factors

Nature Biotechnology volume 21, pages 1208–1214 (2003)Cite this article

A Corrigendum to this article was published on 01 April 2004

Abstract

We have developed a method in which randomized libraries of zinc finger–containing artificial transcription factors are used to induce phenotypic variations in yeast and mammalian cells. By linking multiple zinc-finger domains together, we constructed more than 100,000 zinc-finger proteins with diverse DNA-binding specificities and fused each of them to either a transcription activation or repression domain. The resulting transcriptional regulatory proteins were expressed individually in cells, and the transfected cells were screened for various phenotypic changes, such as drug resistance, thermotolerance or osmotolerance in yeast, and differentiation in mammalian cells. Genes associated with the selected phenotypes were also identified. Our results show that randomized libraries of artificial transcription factors are useful tools for functional genomics and phenotypic engineering.

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Figure 1: Schematic representation of the ZFP library approach.
Figure 2: Various phenotypic changes in yeast induced by artificial ZFP-TFs.
Figure 3: Characterization of ketoconazole-resistant transformants.
Figure 4: Identification of target gene associated with ketoconazole resistance.
Figure 5: Phenotypic changes induced by ZFP-TFs in mammalian cell culture systems.

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Acknowledgements

We thank K.H. Bae, H.C. Shin and J.W. Park for helpful discussions. We also thank Hyun-Mo Ryoo for providing materials, Jae-Ran Lee and Eunjoon Kim for help with immunofluorescence microscopy and K. LaMarco for carefully reading our manuscript. This work was partially supported by the National Research Laboratory Program (M1-0104-00-0048) and by the 21C Frontier Microbial Genomics and Applications Program (MG02-0302-007-2-1-0) of the Korean Ministry of Science and Technology.

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  1. Kyung-Soon Park and Dong-ki Lee: These authors contributed equally to this work.

Authors and Affiliations

  1. ToolGen, Inc., 461-6 Jeonmin-Dong, Yuseong-Gu, Daejeon, 305-390, South Korea

    Kyung-Soon Park, Dong-ki Lee, Horim Lee, Yangsoon Lee, Young-Soon Jang, Yong Ha Kim, Hyo-Young Yang, Sung-Il Lee, Wongi Seol & Jin-Soo Kim

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Correspondence to Jin-Soo Kim.

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The research described in the paper was partially funded by ToolGen, Inc., a privately-held company, and most of the authors are employees. J.-S.K. was one of the founders of the company and is a major shareholder.

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Park, KS., Lee, Dk., Lee, H. et al. Phenotypic alteration of eukaryotic cells using randomized libraries of artificial transcription factors. Nat Biotechnol 21, 1208–1214 (2003). https://doi.org/10.1038/nbt868

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